KR20060013509A - Methods for preventing cattle reproductive diseases - Google Patents

Abstract

The present invention relates to methods for treating or preventing diseases or disorders in a pregnant cow and calf nursing a pregnant cow caused by infection by Bovine Viral Diarrhea Virus (BVDV) Types 1 and 2, Bovine Herpes Virus Type-1 (BHV-1), Bovine Respiratory Syncytial Virus (BRSV), Parainfluenza Virus (PIV3), Campylobacter fetus, Leptospira canicola, Leptospira grippotyphosa, Leptospira hardj-prajitno, Leptospira icterohaemmorrhagiae, Leptospira hardjo-bovis and Leptospira pomona by adminstering to the animal an effective amount of a safe modified live viral combination vaccine further combined with a multivalent bacterin vaccine.

Description

Prevention method of bovine genital disease {METHODS FOR PREVENTING CATTLE REPRODUCTIVE DISEASES}

The present invention provides bovine viral diarrhea virus (BVDV) types 1 and 2, bovine herpes virus type 1 (BHV-1), bovine respiratory syncytial virus (BRSV) by administering an effective amount of a combination vaccine to the animal. , parainfluenza virus type ₃ (PIV 3), Campylobacter page Tooth (Campylobacter fetus), leptospira car Nicola (leptospira canicola), leptospira grip Forty Posadas (leptospira grippotyphosa), leptospira Borg Peter seniyi Har Jo-infrastructure support teuno (leptospira borgpetersenii hardjo-prajitno), leptospira wing terrorism to Mora jiae (leptospira icterohaemmorrhagiae), leptospira Borg Peter seniyi Har trillion - a heritage caused by infection of the Vorbis (leptospira borgpetersenii hardjo-bovis) and leptospira interrogating elegans Pomona (leptospira interrogans pomona) may be given to pregnant cows and calves to prevent diseases or disorders such as abortion. It is about safe vaccination of pregnant pregnant cows. The method of the invention utilizes a combination vaccine, which may be a live preparation that is fully or partially cell inactivated and / or modified.

Five viruses associated with bovine respiratory disease (BRD) complexes, bovine herpes virus type 1 (BHV-1), bovine viral diarrhea virus (BVDV) types 1 and 2, also known as infectious bovine rhinitis virus (IBRV) , Bovine Respiratory Syndrome Virus (BRSV), and Parainfluenza Virus Type 3 (PIV ₃ ) cause respiratory and genital lineage infections that are economically significant for the beef and dairy industry worldwide. BRD causes a wide range of clinical syndromes, including acute onset respiratory disease and miscarriage. BRD in respiratory form is characterized by inflammation, swelling, bleeding and necrosis of the airway mucosa and may involve high fever, anorexia, depression, runny nose, difficulty breathing, and mouth inflammation. Abortion induced by IBRV and BVDV viruses can occur in both third trimester of pregnancy, but mainly occurs during the second trimester of pregnancy and often occurs without other clinical signs. Ellis et al. (1996) JAVMA 208: 393-400; Ellsworth et al. (1994) In: Proceedings, 74 ^th Conference of Research Workers in Animal Disease: 34.

Bovine herpes virus type 1 (BHV-1) is a member of the alphaherpesviridae subfamily and causes various clinical forms of disease in cattle, including respiratory and genital infections, conjunctivitis, encephalitis and miscarriage. do. Existing attempts to modulate BHV-1 infection have been described by the live attenuated virus [Gerber, J. D., et al., 1978, Am. J. Vet. Res. 39: 753-760; Mitchell, D., 1974, Can. Vet. Jour. 15: 148-151], inactivated virus [Frerichs, G. N., et al., 1982, Vet. Rec. 111: 116-122, and one of three major BHV-1 glycoproteins that have been termed gI, gIII, and gIV in the art, for example, Babiuk, LA, et al., 1987, Virology 159: 57-66; van Drunen, S., et al., 1993, Vaccine 11: 25-35]. In addition, the ability of recombinant truncated variants of the BHV-1 gIV glycoprotein to induce mucosal immunity against BHV-1 has been demonstrated. Van Drunen, S., et al., 1994, Vaccine, 12 : 1295-1302]. However, modified-live BHV-1 vaccines recognized in the art are contraindicated in administration to a seropositive or seronegative pregnant cow and also contraindicated to a pregnant cow lactating a calf.

BVDV types 1 and 2 are associated with various clinical syndromes. The virus causes severe primary respiratory disease; Persistently infected (PI) livestock is a major source of infection for susceptible calves; Research has shown that BVDV infects leukocyte-bearing hosts, causing serious and widespread deficiencies in the immune system. Ellis et al. (1996) JAVMA 208: 393-400; Baum et al. (1993) The Compendium Collection: Infectious Disease in Food Animal Practice. Trenton, NJ Veterinary Learning Systems-113-121; Meyling et al. (1987) Agric Pestivirus Infect Rumin 225-231. Abortion or mummification can occur when a pregnant cow is infected, especially during the first trimester of pregnancy. Bolin et al. (1989) Am J. Vet Res 52: 1033-1037. Mucosal disease, another lethal sign of bovine viral diarrhea (BVD), is premature fetal infection with a non-cytoplasmic BVDV biotype, the development of immune tolerance to the virus, and the birth of persistently infected (PI) calves. And subsequent subsequent infection with cytopathic BVDV biotypes. Bolin et al. (1989) Am J. Vet Res 52: 1033-1037. BVDV type 2, most of which has been recognized primarily as a hemorrhagic BVDV isolate in dairy herds, has become a major strain isolated from both BVDV-associated abortion and respiratory cases throughout the United States. Van Oirschot et al. (1999) Vet Micro 64: 169-183.

BVDV is classified into the genus pesitvirus and the Flaviviridae family. This is closely related to the virus that causes border disease in sheep and traditional swine fever. Infected livestock exhibits "mucosal disease" characterized by high temperature, diarrhea, cough and digestive mucosal ulcers. Olafson, et al., Cornell Vet. 36: 205-213 (1946); Ramsey, et al., North Am. Vet. 34: 629-633 (1953). BVD virus can penetrate the placenta of pregnant cows and can result in PI calf birth. Malmquist, J. Am. Vet. Med. Assoc. 152: 763-768 (1968); Ross, et al., J. Am. Vet. Med. Assoc. 188: 618-619 (1986). These calves are immunoresistant to the virus and continue to develop viremia for the rest of their lives. These are potential sources of mucosal disease and are described in Liess, et al., Dtsch. Tieraerztl. Wschr. 81: 481-487 (1974)], the predisposition to infect microorganisms causing diseases such as pneumonia or intestinal disease is high [Barber, et al., Vet. Rec. 117: 459-464 (1985).

According to the findings of BVDV virus growth in cultured cells, two viral biotypes were classified into viruses that induce cytopathic effects in infected cells (cp) and viruses that do not induce cytopathic effects (ncp). See Lee et al., Am. J. Vet. Res. 18: 952-953; Gillespie et al., Cornell Vet. 50: 73-79, 1960. Cp variants may be derived from PI animals pre-infected with ncp virus. Howard et al., Vet. Microbiol. 13: 361-369, 1987; Corapi et al., J. Virol. 62: 2823-2827, 1988. Based on the genetic diversity of the 5 'non-translated region (NTR) and the antigenic differences in the virion surface glycoprotein E2 of the BVD virus, two major genotypes, type 1 and 2, have been proposed. BVDV type 1 is a traditional viral strain that normally causes only minor diarrhea in immunocompetent animals, whereas BVDV type 2 is a highly toxic neoplasm that can cause thrombocytopenia, bleeding and acute fatal disease Virus. Corapi et al., J. Virol. 63: 3934-3943; Bolin et al., Am. J. Vet. Res. 53: 2157-2163; Pellerin et al., Virology 203: 260-268, 1994; Ridpath et al., Virology 205: 66-74, 1994; Carman et al., J. Vet. Diagn. Invest. 10: 27-35, 1998]. Type 1 and 2 BVDV viruses have distinct antigenicity determined by cross-neutralization using panels of monoclonal antibodies (Mabs) and virus-specific antisera generated in animals. Corapi et al., Am. J. Vet. Res. 51: 1388-1394, 1990. Viruses of either genotype may exist as one of two biotypes, cp or ncp virus.

Studies of BVD virus infected animals suggest that BVD virus induces both B-cell and T-cell responses in animals. Donis et al., Virology 158: 168-173, 1987; Larsson et al., Vet. Microbiol. 31: 317-325, 1992; Howard et al., Vet. Immunol. Immunopathol. 32: 303-314, 1992; Lambot et al., J. Gen. Virol. 78: 1041-1047, 1997; Beer et al., Vet. Microbiology. 58: 9-22, 1997.

Numerous BVDV vaccines have been developed using chemically inactivated BVD viral isolates. See Fernelius et al., Am. J. Vet. Res. 33: 1421-1431, 1972; Kolar et al., Am. J. Vet. Res. 33: 1415-1420, 1972; McClurkin et al., Arch. Virol. 58: 119, 1978. Several doses of inactivated viral vaccine are necessary to achieve primary immunity. Some inactivated BVDV vaccines only prevent infection by type 1 BVDV. Beer et al., Vet. Microbiology. 77: 195-208, 2000. Due to the short duration of immunity and inefficient cross protection, fetal protection could not be achieved with inactivated BVDV vaccines. Bolin, Vet. Clin. North am. Food Anim. Pract. 11: 615-625, 1995.

On the other hand, modified-viral virus (MLV) vaccines provide a higher level of protection. Currently, licensed BVDV MLV vaccines use attenuated viruses obtained by repeated passages in cell culture or by Coggins et al., Cornell Vet. 51: 539-, 1961; Philips et al., Am. J. Vet. Res. 36: 135-, 1975], or using chemically modified viruses that exhibit a temperature-sensitive phenotype [Lobmann et al., Am. J. Vet. Res. 45: 2498-, 1984; 47: 557-561, 1986]. A single dose of the MLV vaccine is sufficient to immunize and the duration of immunity can last for years in vaccinated cattle. However, since these vaccines were developed using type I BVDV virus strains, they show the greatest protection against type I viruses. Moreover, the available modified-live BVDV vaccine was not directed to pregnant or lactating pregnant cows.

PIV3 viruses typically cause only minor diseases when acting alone, but they make the airways more susceptible to secondary pathogenic organisms, including IBRV viruses, BRSV, and BVDV, resulting in traditional transport fever (transporting livestock). Fever symptoms that occur during the course of the disease appear. Of the various viruses known to cause respiratory diseases in cattle, the PIV3 virus is the most widely known. Ellis et al. (1996) JAVMA 208: 393-400.

BRSV favors the lower airways, and the depth of infection is largely determined by the immune system's response to viral core proteins. Bolin et al. (1990) Am J Vet Res 51: 703. Diseased cows generally exhibit nonspecific signs, including serous runny nose and tears, often biphasic fever, and frequent dry cough. More severe cows may have a rough cough, difficulty breathing, produce foamy saliva around their mouths, and stop eating and drinking. Ellis et al. (1996) JAVMA 208: 393-400.

Leptospirosis caused by spirochetes of the genus Leptospira is an economically important livestock infection of livestock. Leptospira borgpetersenii serovar hardjo ( L. hardjo ) and L. leptospira borgpetersensei . Interrogans serovar pomona ( L. pomona ) is the two serotypes most commonly associated with bovine leptospirosis worldwide. In one US study of livestock, 29% were L. a. It reacted serologically with Harzo and 23% was L. a. It reacted serologically with pomona. Leptospirosis invades the body through mucous membranes or injured skin and is transmitted through blood. They show directivity to the kidneys and genitals and are usually less directing to the vitreous fluid and the central nervous system of the eye. The most common means of infection is by direct or indirect contact with infected urine, milk or placental fluid, but sexually transmitted diseases and trans-ovarian transmission are also known. Leptospiral infection of cattle can cause acute fever, agaractia, miscarriage, or the birth of immature and weakly infected calves, and can cause reproductive failure and low conception rates. Infections can be treated with antibiotics, but they may be invisible in livestock that are not lactating or pregnant. In these livestock, acute or chronic kidney infections can stick, causing toxic organisms to flow into the urine, which can eventually infect other animals or animal managers. Immunity to leptospira is serotype specific, and vaccines have been available for many years, but most are weak and only induce short-term immunity.

Thus, safe live vaccination of pregnant livestock; There is a need for a method of safe prophylaxis against immunization of various antigens and from propagation from pregnant cattle to lactating offspring. It is also desirable to treat and prevent major causes of infection of respiratory and genital diseases in animals such as cows and calves.

Summary of the Invention

The present invention comprises administering an effective amount of a combination vaccine to a given animal, BVDV type 1 or type 2, BHV-1, PIV ₃ , BRSV, Campylobacter petus, Leptospira canicola, Leptospira grippotifosa, Leptospira borg A method of treating or preventing an animal's disease or disorder caused by infection with one or more of Petersenyi harzo-prasitno, Leptospira ecterohaemorazia, Leptospira borg petersenyi harzo-bovis, and Leptospira interogans pomona to provide.

The method of the invention particularly provides for the protection of animals, eg cattle, from systemic infections and genital diseases. The methods of the present invention provide for the protection of animals such as pregnant cows from miscarriage caused by BHV-1 (IBRV) and BVDV, and persistent fetal infections caused by BVDV types 1 and 2. The methods of the present invention also provide for the protection of animals, such as lactating cows and calf-fed pregnant cows, from infections caused by BVDV types 1 and 2. The methods of the invention also provide for the protection of animals of breeding age, pregnant and lactating animals. Thus, the methods of the present invention provide for the protection of animals prior to breeding and during the conception period. In addition, the methods of the present invention provide for the protection of animals with an annual vaccination history against diseases or infections caused by IBRV, BVDV, PIV3, BRSV, Campylobacter petus and / or Leptospirae . .

Combination vaccines used in the methods of the invention may be complete or partial cellular preparations and / or modified live preparations. Monovalent or combination vaccines administered in accordance with the invention may comprise additional components such as an adjuvant and any second or more antigen. Second antigens are bovine herpes virus type 1 (BHV-1), bovine viral diarrhea virus (BVDV) type 1 or 2, bovine respiratory syncytial virus (BRSV), parainfluenza virus type 3 (PIV ₃ ), leptospiraca Nicola, Leptospira Grippotifosa, Leptospira Borgpetersenini Harzo-Prazitno, Leptospira Ichterohaemorazia, Leptospira Interogans Pomona, Leptospira Borgpetersenyi Harzo-bovis, Lepto brapis ( Lepto bratis ) One selected from, including but not limited to, Philobacter fetus.

The present invention provides a method of treating or preventing a disease or disorder in an animal caused by an infection of IBRV, BVDV, PIV ₃ , BRSV, Campylobacter petus and / or leptospira by administering to the animal an effective amount of a combination vaccine. do. In one embodiment, the present invention is directed to pregnant cows and calves to prevent systemic diseases or disorders, such as miscarriage caused by infection of IBRV, BVDV, PIV ₃ , BRSV, Campylobacter petus and / or leptospiral. Provides a method for immunizing lactating pregnant cows.

In certain embodiments, the vaccines used in the methods of the invention include modified live vaccines and pharmaceutically acceptable carriers, or modified live vaccines and adjuvants.

For purposes of clarity and without limitation, the detailed description of the invention is divided into the following subsections that describe or illustrate particular features, aspects, or applications of the invention.

Definition and abbreviations

As used herein, the term “treatment or prevention” in connection with a disease or disorder refers to toxic BVDV types 1 and 2; IBRV; PIV ₃ ; BRSV; Campylobacteria ; And / or lowering or eliminating the risk of infection by the leptospira antigen, alleviating or alleviating the symptoms of the infection, or promoting recovery from the infection. Treatment is considered therapeutic if there is a decrease in viral or bacterial load, a decrease in lung infection, a decrease in rectal temperature and / or an increase in food intake and / or growth. Treatment is also considered therapeutic if there is a reduction in fetal infection and urinary shedding, for example due to infection with leptospira serotypes Harzo and Pomona. "Infection" means the manifestation of systemic symptoms, including, but not limited to, increased rectal temperature, food intake and / or decreased growth. Infections should also be understood to include respiratory, intestinal, lung and genital subtypes.

The present invention is effective for preventing or lowering rectal temperature and preventing, for example, lactic acid induced by IBRV and infection caused by BVDV types 1 and 2. Thus, the present invention contemplates providing fetal protection from bovine herpes and bovine pestivirus as well as fetal protection from BHV-1 (IBRV) and fetal protection from infections caused by BVDV types 1 and 2. The present invention also contemplates providing protection from persistent fetal infections, eg, persistent BVDV infection. A "persistent fetal infection" refers to active BVDV replication and proliferation that is immunoresistant to BVDV and frequently occurs at high rates for months or years, resulting in the birth of an animal that acts as a permanent source of BVDV in the population. Refers to an infection that occurs during the early fetal development (eg, 45 to 125 days of conception). Such persistently infected animals are also at risk of developing fatal mucosal disease if they have been infected with the cytopathic virus biotype.

The term "combined vaccine" means a divalent or multivalent combination of antigens including modified live antigens. According to the present invention, the combination vaccine comprises modified live infectious IBRV, PIV ₃ , BRSV and BVDV types 1 and 2, Leptospira canicola, Leptospira grippotifossa, Leptospira Borgpetersenyi harzo-pragitno, Leptospira ichterohaemora One or more antigens, veterinary acceptable carriers, and adjuvants, including but not limited to, GiA, Leptospira interogans pomona, Leptospira borgfertersenyi harzo-bovis, Leptospira bratislava and Campylobacter petus can do. In one embodiment the modified live IBRV component is not a temperature sensitive IBRVV. In a preferred embodiment, the BVDV type 1 component exhibits cytopathic effect (cpBVD-1 strain NADL-National Animal Disease Center, United States, Dep. Of Agriculture, Ames, lowa). In another preferred embodiment, the BVDV Type 2 component shows a modified live cytopathic effect (cp BVD-2 strain 53637, ATCC No. PTA-4859). In another preferred embodiment, the modified live antigen is dried, lyophilized or vitrified.

FP™ 4+VL5 또는 Bovi-Shield

FP™5 (화이자, 인크)이다.According to the present invention, the combination vaccine is modified live BHV-1, BVDV types 1 and 2, BRSV, PIV ₃ , Leptospira canicola, Leptospira grippotifosa, Leptospira Borgpetersenyi harzo-prasitno, Leptospira terotero One or more antigens, veterinary acceptable carriers, and any including, but not limited to, Moragea, Leptospira interogans pomona, Leptospira borgfertersenyi harzo-bovis, Leptospira bratislava, and Campylobacter petus It may contain an adjuvant. As used herein, the term “combined vaccine” also refers to one or more modified live antigens, one or more second antigens, and an adjuvant (adjuvant is optional) that prevents or reduces the risk of infection and / or alleviates symptoms of infection. Or multicomponent composition). In a preferred embodiment, the source of the combination vaccine is Bovi-Shield

FP ™ 4 + VL5 or Bovi-Shield

FP ™ 5 (Pfizer, Inc.).

The protective effect of a combination vaccine composition against a pathogen is usually achieved by inducing a combination immune response of a cell-mediated immune response, a humoral immune response, or both in a subject. Generally speaking, the reduction or eradication of BVDV, IBRV, and / or PIV3 infection rates, alleviation of symptoms, or accelerated elimination of the virus from an infected subject suggests a protective effect of the combination vaccine composition. The methods of the present invention are directed to infections caused by BHV-1 (IBRV) or to respiratory infections induced by PIV3 and BRSV, as well as to infections caused by either or both type 1 and type 2 BVD viruses. Provides protection for

The method of the present invention for treating or preventing a disease or disorder in an animal caused by infection by IBRV, BVDV, PIV3, BRSV, Campylobacter petus and / or Leptospira by administering a combination vaccine, is herein prevented. Also referred to as inoculation method.

The term “combination vaccine” that can be used in the methods of the invention includes, for example, inactivated complete or partial seed. C. fetus and / or leptospira cell preparations, modified live BVDV types 1 and 2, and / or one or more modified live antigens such as BHV-1, PIV3 and / or BRSV.

In one embodiment, the vaccine composition of the present invention comprises an effective amount of one or more of the aforementioned BVDV viruses, preferably cpBVD-1 strain NADL (cpBDV-1 strain NADL-National Animal Disease Center, Ames, Iowa); cpBVD-2 strain 53637 (ATCC No. PTA-4859), IBRV strain C-13 (Cutter Laboratories); PIV3 strain Reisinger (University of Nebraska); BRSV strain 375 (Veterinary Research Institute, Iowa Ames). Purified BVDV virus may be used directly in the vaccine composition, or preferably, the BVD virus may be further modified via serial passage in vitro. Typically, the vaccine contains from about 1 x 10 ² to about 1 x 10 ¹⁰ plaque forming or TCID ₅₀ units of virus with a veterinary acceptable carrier and any adjuvant, from 0.1 to 5 ml, preferably about 2 ml It contains in volume. The exact amount of virus in the vaccine composition effective to provide a protective effect can be determined by a professional veterinarian. Suitable veterinary acceptable carriers for use in the vaccine composition can be any of those described below.

In another embodiment, the vaccine composition of the present invention comprises an effective amount of at least one of the aforementioned BHV-1 (IBRV) viruses, preferably IBRV strain C-13 (Cutter Laboratories), typically the vaccine is about Virus from 1 x 10 ² to about 1 x 10 ¹⁰ plaque or colony forming unit is contained in a volume of 0.1 to 5 ml, preferably about 2 ml, together with a veterinary acceptable carrier and adjuvant. The exact amount of virus in the vaccine composition effective to provide a protective effect can be determined by a professional veterinarian. Suitable veterinary acceptable carriers for use in the vaccine composition may be any of those described below. Typical routes of administration are intramuscular or subcutaneous injections of about 0.1 to about 5 ml of vaccine. Vaccine compositions of the present invention may contain additional active ingredients, such as other vaccine compositions for BVDV, for example WO 95/12682, WO 99/55366, US Pat. No. 6,060,457, US Pat. No. 6,015,795, US Pat. No. 6,001,613 And US Pat. No. 5,593,873.

Vaccination can be achieved through single or multiple inoculations. If desired, serum may be collected from the animals so inoculated and tested for the presence of antibodies to BVD virus and IBRV virus.

In another aspect of the invention, the vaccine composition is used to treat BVDV infection. Accordingly, the present invention provides a method of treating an infection in an animal subject caused by BVD virus type 1 or type 2, or a combination of type 1 and type 2, by administering to the animal a therapeutically effective amount of the BVD virus of the present invention. to provide. In another embodiment, the vaccine compositions of the present invention are effective for improving population fertility and reducing the risk of disease transmission from livestock to humans.

By “animal subject” is meant to include all animals susceptible to BVDV, BHV, PIV3, BRSV or Leptospira infections, such as cattle, sheep and pigs. In one embodiment, the animal subject is a cow, calf or heifer that is naïve. In a preferred embodiment, the animal subject is a lactating cow. In another preferred embodiment, the animal subject is a pre-breeding cow, a pregnant cow or a pregnant cow feeding a calf.

In practicing the methods of the invention, the vaccine composition of the invention is preferably administered to livestock via an intramuscular or subcutaneous route, but other routes of administration, such as oral, nasal (eg, aerosols or other acupuncture). Administration), lymph node, intradermal, intraperitoneal, rectal or vaginal administration, or a combination route may be used. Intramuscular administration to the neck of the animal is preferred. A booster regimen may be required and the dosage regimen may be adjusted to provide optimal immunity.

"Immunogenic" means the ability of a BVD virus to elicit an immune response in an animal against type 1 or type 2 BVD viruses, or against both type 1 and type 2 BVD viruses. The immune response may be a cellular immune response primarily mediated by cytotoxic T-cells, or a humoral immune response mediated primarily by helper T-cells, which in turn activate B-cells resulting in antibody production. .

According to the present invention, the virus is preferably attenuated by a series of passages in cell culture prior to use in immunogenic compositions. Modification methods are well known to those skilled in the art.

Preferred viruses to be included in the immunogenic composition in the methods of the invention are BVDV cpBVDV NADL (Type 1). Another preferred virus to be included in the immunogenic composition of the present invention is cp53637 (ATCC No. PTA-4859). A further preferred virus to be included in the immunogenic composition of the present invention is IBRV strain C-13. Another preferred virus to be included in the immunogenic compositions of the invention is the PIV3 strain racer. Another preferred virus to be included in the immunogenic composition of the present invention is BRSV strain 375.

Immunogenic compositions used in the methods of the present invention may also contain additional active ingredients, such as other immunogenic compositions against BVDV, for example, pending US patent application Ser. No. 08 / 107,908, WO 95/12682, WO 99 / 55366, US Pat. No. 6,060,457, US Pat. No. 6,015,795, US Pat. No. 6,001,613, and US Pat. No. 5,593,873.

아주반트, 사포닌, 퀼 (Quil) A, QS-21 (캠브리지 바이오텍 인크, 메사추세츠주 캠브리지), GPI-0100 (갈레니카 파마슈티칼스 인크, 앨라배마주 버밍햄) 또는 기타 사포닌 분획, 모노포스포릴 지질 A, 아브리딘 (Avridine) 지질-아민 아주반트, 이. 콜라이 (E. coli)로부터의 열-불안정 장독소 (재조합 기원 또는 그 밖의 기원), 콜레라 독소, 또는 무라밀 디펩티드가 포함되지만, 이에 제한되지 않는다. 본 발명의 면역원성 조성물은 하나 이상의 기타 면역조정제, 예를 들면 인터루킨, 인터페론 또는 기타 사이토카인을 추가로 포함할 수 있다. 본 발명의 방법에 이용된 면역원성 조성물은 젠타마이신 및 메르티올레이트를 포함할 수도 있다. 본 발명의 맥락에서 유용한 아주반트와 첨가제의 양 및 농도는 당업자에 의해 용이하게 결정될 수 있지만, 본 발명은 백신 조성물 2 ml 용량 당 약 50 ㎍ 내지 약 2000 ㎍의 아주반트, 바람직하게는 약 500 ㎍의 아주반트를 포함하는 조성물을 고려한다. 또 다른 바람직한 양태에서, 본 발명은 약 1 ㎍/ml 내지 약 60 ㎍/ml의 항생제, 보다 바람직하게는 약 30 ㎍/ml 미만의 항생제를 포함하는 백신 조성물을 고려한다.In addition, the immunogenic and vaccine compositions used in the methods of the invention may comprise one or more veterinary acceptable carriers. "Vetically acceptable carrier" as used herein includes all solvents, dispersion media, coatings, adjuvants, stabilizers, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, adsorption delaying agents, and the like. Diluents may include water, saline, dextrose, ethanol, glycerol, and the like. Isotonic agents can include sodium chloride, dextrose, mannitol, sorbitol, lactose, and the like. Stabilizers include albumin and the like. Adjuvant includes RIBI adjuvant system (Ribi Inc.), alum, aluminum hydroxide gel, cholesterol, oil-in-water emulsion, water-in-oil emulsion, e.g. Freund's complete and incomplete adjuvant, block copolymer (CytRx, Atlanta GA) , SAF-M (Chiron, Emeryville, CA), AMPHIGEN

Adjuvant, saponin, quil A, QS-21 (Cambridge Biotech Inc., Cambridge, Mass.), GPI-0100 (Galenica Pharmaceuticals Inc, Birmingham, Alabama) or other saponin fraction, monophosphoryl lipid A, Avridine lipid-amine adjuvant, E. Heat-labile enterotoxin (recombinant origin or other origin), cholera toxin, or muramyl dipeptides from E. coli . The immunogenic compositions of the invention may further comprise one or more other immunomodulators, such as interleukin, interferon or other cytokines. Immunogenic compositions used in the methods of the present invention may comprise gentamicin and merthioleate. Amounts and concentrations of adjuvants and additives useful in the context of the present invention can be readily determined by one skilled in the art, but the present invention provides from about 50 μg to about 2000 μg of adjuvant, preferably about 500 μg per 2 ml dose of vaccine composition. Consider a composition comprising an adjuvant of. In another preferred embodiment, the present invention contemplates a vaccine composition comprising about 1 μg / ml to about 60 μg / ml antibiotic, more preferably less than about 30 μg / ml antibiotic.

Immunogenic compositions used in the methods of the invention can be prepared in various forms depending on the route of administration. For example, the immunogenic compositions of the invention may be in the form of sterile aqueous solvents or suspensions suitable for injection, or in lyophilized form using freeze-drying techniques. Lyophilized immunogenic compositions are typically maintained at about 4 ° C. and can be reconstituted in stable solutions, such as saline and / or HEPES, with or without adjuvant.

The immunogenic compositions of the invention can be administered to animal subjects to induce an immune response against type 1 or type 2 BVD viruses, or against both type 1 and type 2 BVD viruses. Accordingly, another aspect of the invention provides an immune response against type 1 or type 2 BVD virus, or both type 1 and type 2 BVD virus, by administering to the animal subject an effective amount of the immunogenic composition of the invention mentioned above. Provide a way to stimulate. "Animal subject" is meant to include all animals susceptible to BVDV infection, such as cattle, sheep and pigs.

According to the methods of the invention, preferred immunogenic compositions for administration to an animal subject include BVDV cpNADL virus and / or BVDV cp53637 virus. Immunogenic compositions containing BVDV virus, preferably live virus modified by serial passage in culture, are preferably administered to livestock via intramuscular or subcutaneous routes, but other routes of administration, for example, It may also be administered by oral, nasal, lymph node, intradermal, intraperitoneal, rectal or vaginal administration, or by a combination route.

Immunization protocols can be optimized using procedures well known in the art. A single dose may be administered to the animal, or two or more inoculations may be performed at 2-10 week intervals. Depending on the age of the animal, the immunogenic or vaccine composition may be re-administered. For example, the present invention contemplates vaccination of healthy livestock six months before birth and then again vaccination at six months of age. In another example, the present invention is directed to pre-breeding livestock about 5 weeks before breeding (or before joining a herd) and optionally re-vaccinated about 2 weeks prior to breeding or during conception, thereby providing BVDV type 1 and Consider protecting the fetus from infection caused by 2. A single dose of a composition of the present invention may be administered about three to four weeks after the first dose. Re-vaccination of a single dose of the combination vaccine of the present invention every six months is also contemplated to prevent BVDV fetal infection.

The degree and nature of the immune response induced in livestock can be assessed using various techniques. For example, serum can be collected from inoculated animals and tested for the presence of specific antibodies against BVDV virus, for example in conventional virus neutralization assays.

As used herein, the term “cow” refers to bovine animals, including but not limited to (caster) bulls, bulls (not castrated), unborn cows, cows, and calves. Cows as used herein refers to pregnant and lactating bovine animals. Preferably, the methods of the present invention apply to non-human mammalian animals, preferably lactating or pregnant cows and their fetuses, or lactating calves.

The term "therapeutically effective amount" or "effective amount" refers to an amount of the combination vaccine sufficient to elicit an immune response in the animal to which the combination vaccine is administered. Immune responses may include, but are not limited to, induction of cellular and / or humoral immunity. The amount of therapeutically effective vaccine may vary depending on the particular virus used, the condition of the cow and / or the extent of infection and may be determined by the veterinarian.

Inactivation (partial or whole cell) and modified live vaccine

Inactivated or modified live vaccines for use in the methods of the invention can be prepared using a variety of methods known in the art.

For example, BVDV isolates can be obtained directly from the uterus of infected cows using known techniques.

BVDV isolates can be attenuated using a variety of known methods, including, for example, serial passage. In addition to modified live viral isolates, the vaccine product used in the methods of the present invention may also contain an appropriate amount of one or more adjuvants that are commonly used. Suitable adjuvants include mineral gels such as aluminum hydroxide; Surface active substances such as lyso lecithin; Glycosides such as saponin derivatives such as quill A or GPI-0100; Polyhydric alcohols; Polyvalent anions; Non-ionic block polymers such as Pluronic F-127 (B.A.S.F., USA); Peptides; Inorganic oils such as Montanide ISA-50 (Sepic, Paris, France), Carbopol, Amphigen, Amphigen Mark II (Hydronics, USA), Alhydrogel Emulsions of oils, for example inorganic oils (eg BayolF / Arlacel A) and water, or emulsions of vegetable oils, water and emulsifiers (eg lecithin); alum; Bovine cytokine; cholesterol; And combinations of adjuvants may be included, but are not limited thereto. In a preferred embodiment, saponin-containing oil-in-water emulsions are usually microfluidized.

FP™ 4+VL5 (화이자, 인크)이다. 본 발명의 방법에 사용하는데 바람직한 BVDV 유형 2의 공급원은 cp BVDV 균주 53637 (Univ. Guelph, Guelph, Ont.) (ATCC No. PTA-4859)이다.A particularly preferred source of BVDV type 1 for use in the methods of the present invention is Bovi-Shield containing BVDV strain NADL (National Animal Disease Center (NADC), USDA, Ames, Iowa).

FP ™ 4 + VL5 (Pfizer, Inc.). A preferred source of BVDV type 2 for use in the methods of the invention is cp BVDV strain 53637 (Univ. Guelph, Guelph, Ont.) (ATCC No. PTA-4859).

Preferably, strains NADL and 53637 are modified live strains. According to the invention, the strain of the invention can be supplemented with a commercial adjuvant, preferably quill A-cholesterol-ampygen (hydronix, USA). Preferred doses of the immunogenic and vaccine compositions of the invention are about 2.0 ml. Preservatives may be included in the compositions used in the methods of the invention. Preservatives contemplated by the present invention include gentamicin and merthioleate. A carrier, preferably PBS, may also be added. For example, it is known in the art to produce modified live vaccines that attenuate toxic strains by passage in culture.

Modified live BVDV isolates include bovine herpes virus type 1 (BHV-1), bovine respiratory syncytial virus (BRSV), parainfluenza virus (PIV3), campylobacter petus, leptospira canicola, leptospira grippotifosa, leptospira It can also be combined with bacteria and viruses, including but not limited to, borgpetersenyi harzo-prasitno, leptospira ecterohaemoria, leptospira borgfertersenyi harzo-bovis, and leptospira interogans pomona.

Dosage and Mode

According to the present invention, an effective amount of a combination vaccine administered to a livestock, including pregnant and lactating pregnant cows, is directed against fetal infections and diseases associated with BHV-1 and bovine viral diarrhea viruses (types 1 and 2). Provides valid immunity In one embodiment, the combination vaccine is administered to the calf in two doses about 3-4 weeks apart. For example, the first administration is performed about 1 to about 3 months of age. The second administration is carried out with the combination vaccine about 1 to about 4 weeks after the first administration.

In a preferred embodiment, the first administration, eg pre-vaccination, is carried out about 4-5 weeks prior to animal breeding. The second administration is carried out about 3-4 weeks after animal breeding. Administration of subsequent vaccine doses is preferably performed on an annual basis. In another preferred embodiment, animals vaccinated prior to 6 months of age should be re-vaccinated after 6 months of age. Although administration of subsequent vaccine doses is preferably performed on an annual basis, administration of subsequent vaccine doses on a two year and six month basis is also contemplated by the present invention.

The amount of effective combination vaccine is determined by the components of the vaccine and the schedule of administration. Typically, when a modified bovine viral diarrhea virus preparation is used in a vaccine, BVDV (from about 10 ² to about 10 ¹⁰ TCID ₅₀ units, preferably from about 10 ⁵ to about 10 ⁸ TCID ₅₀ units per dose) ( The amount of vaccine containing types 1 and 2) is effective when administered twice to animals for about 3 to 10 weeks. Preferably, when administered twice to an animal for about 3 to 10 weeks, the combination vaccine that provides effective immunization is about 10 ⁵ to about 10 ⁸ TCID ₅₀ units per dose, more preferably about 10 ⁶ TCID _It contains ₅₀ units of BVDV (types 1 and 2). First administration is performed about 5 weeks prior to animal breeding. The second administration is carried out about 3-4 weeks after animal breeding. Administration of subsequent vaccine doses is preferably performed on an annual basis. Animals vaccinated prior to 6 months of age should be re-vaccinated after 6 months of age. Administration of subsequent vaccine doses is preferably performed on an annual basis.

FP™ 4+VL5 (화이자, 인크)를 투여하는 경우에는, 이러한 생성물을 2회 투여하는 것이 바람직한데, 매회 투여시의 양은 약 0.1 내지 약 5.0 ml, 바람직하게는 약 1.5 내지 약 2.5 ml, 보다 바람직하게는 약 2 ml이다. 제1 투여는 동물 번식시키기 약 5주 전에 수행한다. 제2 투여는 동물 번식 후 약 3 내지 4주째에 수행한다. 후속 백신 용량의 투여는 연간 기준하여 수행하는 것이 바람직하다. 생후 6개월 이전에 예방접종한 동물은 생후 6개월 이후에 재예방접종해야 한다. 후속 백신 용량의 투여는 연간 기준하여 수행하는 것이 바람직하다.According to the invention, the preferred product, Bovi-Shield

When administering FP ™ 4 + VL5 (Pfizer, Inc.), it is preferred to administer these products twice, each amount being about 0.1 to about 5.0 ml, preferably about 1.5 to about 2.5 ml, more Preferably about 2 ml. The first administration is carried out about five weeks before animal breeding. The second administration is carried out about 3-4 weeks after animal breeding. Administration of subsequent vaccine doses is preferably performed on an annual basis. Animals vaccinated prior to 6 months of age should be re-vaccinated after 6 months of age. Administration of subsequent vaccine doses is preferably performed on an annual basis.

According to the present invention, administration can be accomplished by known routes including oral, intranasal, topical, transdermal, and parenteral (eg, intravenous, intraperitoneal, intradermal, subcutaneous or intramuscular). Preferred routes of administration are subcutaneous or intramuscular administration.

The present invention also seeks to eliminate the need to administer additional doses to the calf prior to annual vaccination to produce and / or maintain immunity to infection, followed by a first single dose followed by annual vaccination.

, 알히드로겔, 오일 유제, 예를 들면, 무기 오일 (예를 들면, BayolF/Arlacel A)와 물의 유제, 또는 식물성 오일, 물 및 유화제 (예를 들면, 레시틴)의 유제; 명반; 소 사이토카인; 및 아주반트의 조합물]를 포함할 수 있다.Combination vaccines administered in accordance with the present invention may comprise additional ingredients, such as adjuvant [eg, mineral gels such as aluminum hydroxide; Surface active substances such as cholesterol, lysolecithin; Glycosides such as saponin derivatives such as quill A, QS-21 or GPI-0100; Polyhydric alcohols; Polyvalent anions; Non-ionic block polymers such as Pluronic F-127; Peptides; Inorganic oils, for example Montanide ISA-50, Carbopol, Ampicen

, Alhydrogels, oil emulsions, for example emulsions of inorganic oils (eg BayolF / Arlacel A) with water, or emulsions of vegetable oils, water and emulsifiers (eg lecithin); alum; Bovine cytokine; And combinations of adjuvants].

According to the present invention, administration of an effective amount of a combination vaccine administered to livestock at approximately three months of age provides effective immunity against respiratory infections and genital diseases and lowers miscarriage.

The invention also relates to BVDs (types 1 and 2), IBRV, PIV3, BRSV, Leptospira canicola, Leptospira grippotifosa, Leptospira Borgpetersenyi harzo-prasitno, Leptospira ecterohaemorage, Leptospira intraspira One or more doses, preferably two doses, of the combination vaccine to immunize livestock against infections caused by Pomona, Leptospira Borgpetersensei Harzo-Vorbis, Leptospira Bratislava, and Campylobacter petus By administering to BHV-1 and BVDV (types 1 and 2) and respiratory diseases caused by IBRV, BVDV (types 1 and 2), PIV3, BRSV, campylobacterosis and leptospirosis A method of immunizing livestock, including but not limited to cows, calves and preparative cows of infertility, is provided.

In a preferred embodiment, the vaccine of the present invention is administered subcutaneously. In another preferred embodiment, the vaccine is administered intramuscularly. Moreover, the vaccine dose comprises about 1 to about 7 ml, preferably about 2 ml, each ml preferably containing about 10 ² to about 10 ¹⁰ TCID ₅₀ units of virus per dose. Combination vaccines are desired to be administered twice to animals; It is administered once about 1 to 3 months after birth, and once again after about 1 to 5 weeks. The present invention also contemplates re-vaccination at a single dose on a six month cycle and prior to breeding.

The invention also relates to BVDs (types 1 and 2), IBRV, PIV3, BRSV, Leptospira canicola, Leptospira grippotifosa, Leptospira Borgpetersenyi harzo-prasitno, Leptospira ecterohaemorage, Leptospira intraspira One or more doses, preferably two doses of the combination vaccine to immunize the fetus against infections caused by Pomona, Leptospira Borgpetersensei Harzo-Bovis, Leptospira Bratislava, and Campylobacter petus : A method of protecting a fetus from fetal infection and persistent fetal infection, including administering to a mother animal). Combination vaccines are desired to be administered twice to animals; It is administered once about 5 weeks prior to breeding and once again about 3 to 4 weeks after breeding.

The present invention also contemplates administering an effective amount of a combination vaccine to an animal, preferably a livestock, to treat or prevent disorders, including persistent fetal infections and genital diseases, such as abortions in an animal.

The invention is further illustrated by, but not limited to, the following examples.

4+L5) 제형을, 예방접종하지 않은 임신한 가축과 예방접종한 임신한 가축에게 투여하는 것에 대한 안전성 여부를 평가하기 위해, 집중적인 안전성 연구를 수행하였다. 연구용 동물은 임신한지 대략 160일 내지 220일이 된, 20마리의 예방접종하지 않은 교배종 식용 출산 미경험 암소 (처리 그룹 T1)와 59마리의 미리 예방접종한 교배종 식용 출산 미경험 암소 (처리 그룹 T2=14, T3=15, T4=15, 및 T5=15)로 구성되었다. 예방접종한 동물에 대한 번식 이전의 예방접종 전력은, 번식시키기 대략 150일 전에 T3 및 T5 동물 각각에게 소 비기관염-바이러스 설사-파라인플루엔자₃-호흡기 합포체 변형-생 바이러스 백신 (Bovi-Shield

4)을 투약하고, 모든 T2, T3, T4, 및 T5 동 물에게는 번식시키기 대략 30일 전에 소 비기관염-바이러스 설사-파라인플루엔자₃-호흡기 합포체 변형-생 바이러스 백신/캄필로박터 페투스-렙토스피라 카니콜라-그립포티포사-하르조-익테로해모라지애-포모나 박테린 (PregGuard

10)을 투약하는 것으로 구성되었다.Bovine rhinitis-virus diarrhea-parainfluenza ₃ -respiratory complexes modified-live virus vaccine / Leptospira canicola-gripportifosa-harzo-ecterohaemorrhagia containing 10-fold dose of modified live virus (MLV) component Pomona bacterin (Bovi-Shield

In order to assess the safety of administering 4 + L5) formulations to vaccinated pregnant and vaccinated pregnant livestock, intensive safety studies were performed. The study animals were fed 20 unvaccinated hybrid edible unborn cows (treatment group T1) and 59 pre-vaccinated hybrid edible unborn cows (treatment group T2 = 14), approximately 160-220 days pregnant. , T3 = 15, T4 = 15, and T5 = 15). Vaccination history prior to breeding for vaccinated animals was evaluated in each of the T3 and T5 animals approximately 150 days prior to breeding for bovine nasalitis-virus diarrhea-farinfluenza ₃ -respiratory complex modified-live virus vaccine (Bovi-Shield).

4) Dosage to all T2, T3, T4, and T5 animals about 30 days prior to breeding Bovine rhinitis-virus diarrhea-Parlinefluenza ₃ -Respiratory conjugate modification-Live virus vaccine / campylobacter petus- Leptospira canicola-grip fortyphosa-harzo-ecterohaemorgia-pomona bacterin (PregGuard

10).

4+L5 제형을 투여하였다. 연구용 동물을 대상으로 하여 건강 상태와 분만을 통한 유산에 대해 매일 관찰하고, 예방접종에 대한 혈청학적 반응을 결정하며, 유산 증례에 대한 진단 시험을 수행하고, 건강 상태와 혈청학적 상태를 각 송아지에 대해 결정하였다. 전반적으로, BVDV (9)에 대한 수유 전의 중화 항체를 지니면서 탄생한 송아지 또는 유산아 (6)에 의해 입증된 바와 같이 0일째 예방접종에 의해 T1 중의 15마리의 임신한 가축이 영향을 받았다. 0일째에 IBRV 중화 항체에 대해 음성 (2)이었던 11마리의 T1 출산 미경험 암소 중의 6마리 (55%)에게서 25일 내지 43일째에, IBRV 감염으로 인한 유산이 발생하였다. 나머지 T1 출산 미경험 암소 (14)는 건강한 송아지를 분만하였지만; BVDV 중화 항체를 지니면서 탄생한 송아지는, 0일째에 BVDV 중화 항체에 대해 음성이었던 12마리의 출산 미경험 암소 중의 9마리 (75%)에게서 자궁내 노출이 일어났다.On day 0, intramuscular dose of Bovi-Shield containing 10 doses of MLV component

4 + L5 formulation was administered. Investigate animals daily for health status and miscarriage through labor, determine serological responses to vaccinations, perform diagnostic tests on abortion cases, and monitor health and serological status in each calf. Was determined. Overall, 15 pregnant livestock in T1 were affected by day 0 vaccination, as evidenced by calf or aborted infants (6) born with neutralizing antibodies prior to feeding to BVDV (9). From day 25 to day 43, abortion from IBRV infection occurred in 6 of 55 (55%) of 11 T1 naïve cows that were negative (2) for IBRV neutralizing antibodies on day 0. The remaining T1 naïve cows 14 delivered healthy calves; Calf born with BVDV neutralizing antibodies developed intrauterine exposure in 9 of 75 (75%) naïve cows that were negative for BVDV neutralizing antibodies on day 0.

In comparison, there was no adverse effect on the pregnancy of pre-vaccinated animals in T2, T3, T4, and T5. All of these animals (59/59) delivered healthy calves and 58 of these calves (one calf was fed prior to sample collection) were negative for neutralizing antibodies to IBRV and BVDV.

백신으로 예방접종시킨 출산 미경험 암소의 임신은 10 용량의 MLV 성분을 함유하는 시험용 백신 제형의 투여에 의해 불리한 영향을 받지 않았는데, 이는 이들 성분이 예방접종한 임신한 가축에게서 안전하다는 것을 입증해주었다.Bovi-Shield before breeding

Pregnancy of naïve cows vaccinated with the vaccine was not adversely affected by administration of a test vaccine formulation containing 10 doses of the MLV component, demonstrating that these components are safe in vaccinated pregnant livestock.

research plans:

Treatment number Pre-Research Virus Vaccination History * Number of animals Bovi-Shield 4 + L.5 Vaccination About 150 days before breeding About 30 days before breeding Work IM capacity * T1 none none 20 0 One T2 none Preg-Guard 10 14 0 One T3 Bovi-Shield 4 Preg-Guard 10 15 0 One T4 none Preg-Guard 10 15 0 One T5 Bovi-Shield 4 Preg-Guard 10 15 0 One * Each 2ml IM (Intramuscular) dose was formulated to contain 10-dose MLV components.

process:

Pre-processing Steps:

Animals were selected for this study based on minimizing calf delivery intervals. Pregnant animals with no history of vaccination for IBRV and BVDV were assigned to treatment group T1. Animals were assigned to the treatment groups according to a completely random setting. In summary, the animals without any vaccination history for IBRV and BVDV were negative for BVDV and had serum neutralizing antibody titers for IBRV, and type 1 and 2 BVDV prior to vaccination. Approximately 150 days before breeding, certain doses of Bovi-Shield 4 (Bovine Nasalitis-Virus Diarrhea-Farlinefluenza ₃ -Respiratory Syndrome Modified-Live Virus Vaccine) were administered to T3 and T5 animals, respectively. Approximately 30 days prior to breeding, a dose of Preg-Guard 10 was administered to each animal in the T2 and T3, and T4 and T5 groups. The vaccine in this study contained 37 log ₁₀ TCID ₅₀ of IBRV and 2.7 log ₁₀ TCID ₅₀ of BVDV per 2-ml dose.

Processing steps:

Beginning at day -1, all animals were observed daily for general health status and miscarriage through childbirth. On day 0, pregnancy status of each animal was confirmed and two serum separation tubes (SSTs) of blood were collected from each animal. Bovi-Shield 4 + L5 vaccines were prepared and administered to each of 79 study animals a 2-ml intramuscular dose containing 10-dose of the four MLV components of this vaccine. All animals were observed for adverse reactions for approximately 1 hour after vaccination.

All aborted animals were identified and two SST blood samples (acute) were collected, processed into serum, labeled and stored frozen. Aborted fetal samples available from 5 of 6 aborted infants were shipped to a diagnostic laboratory. After approximately two weeks, two additional SST blood samples (recovery phases) were collected from the mother animals, processed into serum, labeled and stored frozen.

Childbirth Steps:

On calf delivery days, the health and lactation status of each calf was determined and blood samples (2-SSTs) were collected from each calf. Post-feeding blood samples (2-SSTs) were collected from each calf on day 3 of birth. Blood samples collected from calves were processed into serum, labeled, frozen and then tested.

Serum test:

Serum obtained from naïve cows and calves was tested for IBRV, and neutralizing antibodies against type 1 and 2 BVDV. If calf was not observed to be fed prior to sample collection, serum prior to and after lactation was also determined for total serum IgG concentration, whose pre-serum serum was positive for neutralizing antibodies to IBRV and / or BVDV. (≧ 4).

result:

Table 1: Test Vaccine Modified Live Virus Components

Table 2: Geometric Means and Ranges of IBRV Neutralizing Antibody Titers on Days 0 and 28

Table 3: Geometric Means and Ranges of Type 1 BVDV Neutralizing Antibody Titers on Days 0 and 28

Table 4: Geometric Means and Ranges of Type 2 BVDV Neutralizing Antibody Titers on Days 0 and 28

Table 5: Summary of Effects on Pregnancy Due to Day 0 Treatment

Table 6: Summary of Individual Results for Processing Group T1

Vaccine Modified Live Virus Components:

As summarized in Table 1, the Bovi-Shield 4 + L5 formulation administered to each study animal on day 0 was modified live IBRV, BVDV, PIB3, and 7.0, 5.5, 7.5, and 5.7 log ₁₀ TCID ₅₀ of the vaccine. Each contained a BRSV component.

Serological response to vaccination:

Tables 2, 3 and 4 contain geometric mean and range of serum neutralizing antibody titers for IBRV, Type 1 BVDV, and Type 2 BVDV, respectively, in each treatment group before vaccination (day 0) and post vaccination (day 28). Doing.

The mean pre-vaccination IBRV antibody titer was 2 for non-vaccinated (T1) animals and 11 of 20 T1 animals were negative (<2) for neutralizing antibodies to IBRV. The mean pre-vaccination IBRV antibody titers (14, 16, 16, and 21) for vaccinated animals (T2, T3, T4, and T5) are the preimmune vaccination regimen (1 or 2 doses) and the prescription (minimum immunization). Or IBRV component of release level). Mean antibody titers against T1, T2, T3, T4, and T5 increased significantly on average at 86, 130, 94, 106, and 97 in response to vaccination on day 28, respectively. Nineteen of the 20 non-vaccinated animals (T1) were negative for both type 1 and type 2 BVDV antibodies on day 0. Vaccinated (T2, T3, T4, and T5) mean type 1 (223, 215, 315, and 250) and type 2 (44, 57, 84, and 58) BVDV neutralizing antibody titers were compared with the immunization regime It was similar regardless of prescription. On day 28, T1 mean titers for Type 1 (181) and Type 2 (15) BVDV increased significantly, while Type 1 (208, 228, 277, and 231) and Type 2 BVDV (49, 63, 58, And T2, T3, T4, and T5 mean titers for 58) were comparable to pre-vaccination levels.

Effect of treatment on day 0 on pregnancy:

The effect of day 0 treatment (vaccination) on pregnancy in each treatment group is summarized in Table 5. Overall, 15 of T1 pregnant animals were affected by vaccination, as evidenced by calf or aborted infants (6) born with neutralizing antibodies prior to feeding to BVDV (9). In comparison, there was no adverse effect on the pregnancy of pre-vaccinated animals in T2, T3, T4, and T5. All of these animals (59/59) delivered healthy calves and 58 of these calves (one calf was fed prior to sample collection) were negative for neutralizing antibodies to IBRV and BVDV.

Summary of individual results for treatment group T1:

Individual abortion and delivery results for non-vaccinated animals (T1) are summarized in Table 6. From day 25 to day 43, abortions from IBRV infection occurred in 6 of 55 (55%) of 11 T1 naïve cows that were negative (<2) for IBRV neutralizing antibodies on day 0. The remaining T1 naïve cows 14 delivered healthy calves; Calf born with BVDV neutralizing antibodies developed intrauterine exposure in 9 of 75 (75%) naïve cows that were negative for BVDV neutralizing antibodies on day 0.

conclusion:

백신으로 예방접종시킨 출산 미경험 암소의 임신은, 10 용량의 BOVI-SHIELD

의 MLV 성분을 함유하는 시험용 백신 제형을 투여함으로써 불리한 영향을 전혀 받지 않았는데, 이는 이들 바이러스가 예방접종한 임신한 가축에게서 안전하다는 것을 입증해주었다.BOVI-SHIELD prior to breeding

Pregnancy of childbirth inexperienced cow whom we vaccinated with vaccine is BOVI-SHIELD of 10 doses

Administration of a test vaccine formulation containing the MLV component of was not adversely affected, demonstrating that these viruses are safe in vaccinated pregnant livestock.

Vaccine modified live virus components for testing BOVI-SHIELD 4 + L5 Log ₁₀ TCID ₅₀ per 2-ml dose IBRV BVDV PIV ₃ BRSV Trial Vaccine Formulation * 7.0 5.5 7.5 5.7

* Each 2-ml dose of the test vaccine was formulated to contain 10-dose modified live viral components, so each study animal received an additional 1.0 log ₁₀ TCID ₅₀ of each viral component.

Geometric Mean and Range of IBRV Neutralizing Antibody Titers at Days 0 and 28 Processing group Number of immunizations before breeding Number of animals 0 day GMT ** 28 days GMT ** Zero-day titer range 28-day titer range T1 0 (not applicable) 20 2 86 *** <2-16 16-304 T2 1 (IBRV) 14 14 130 7-91 54-431 T3 2 (IBRV) 15 16 94 10-27 38-128 T4 1 (BVDV) 15 16 106 5-181 27-304 T5 2 (BVDV) 15 21 97 8-152 27-431

** A titer of <2 was estimated as 1.0 prior to the log ₂ transformation to determine GMT (geometric mean titer).

*** 11 of 20 T1 animals were negative (<2) for neutralizing antibodies to IBRV.

Geometric Means and Ranges of Type 1 BVDV Neutralizing Antibody Titers on Days 0 and 28 Processing group Number of immunizations before breeding Number of animals 0 day GMT ** 28 days GMT ** Zero-day titer range 28-day titer range T1 0 (not applicable) 20 One 181 *** <2-1218 32-1218 T2 1 (IBRV) 14 223 208 76-1024 54-724 T3 2 (IBRV) 15 215 228 38-861 91-512 T4 1 (BVDV) 15 315 277 76-861 45-724 T5 2 (BVDV) 15 250 231 64-861 64-861

** A titer of <2 was estimated as 1.0 prior to the log ₂ transformation to determine GMT (geometric mean titer).

*** 19 of 20 T1 animals were negative for neutralizing antibodies to type 1 BVDV.

Geometric Means and Ranges of Type 2 BVDV Neutralizing Antibody Titers on Days 0 and 28 Processing group Number of immunizations before breeding Number of animals 0 day GMT ** 28 days GMT ** Zero-day titer range 28-day titer range T1 0 (not applicable) 20 One 15 *** <2-609 3-362 T2 1 (IBRV) 14 44 49 8-256 13-181 T3 2 (IBRV) 15 57 63 4-215 23-128 T4 1 (BVDV) 15 84 58 27-431 13-181 T5 2 (BVDV) 15 58 58 13-181 16-181

** A titer of <2 was estimated as 1.0 prior to the log ₂ transformation to determine GMT (geometric mean titer).

*** 19 of 20 T1 animals were negative for neutralizing antibodies to type 2 BVDV.

Summary of effects on pregnancy due to day 0 treatment Processing group Number of immunizations before breeding Number of animals Miscarriage Healthy Calf Count ** Number of calves positive for VN antibody prior to lactation ** Number of Pregnancies Affected by Day 0 Treatment IBRV BVDV-1 BVDV-2 T1 0 (not applicable) 20  6 14 0 9 7 15 T2 1 (IBRV) 14 0 14 0 0 0 0 T3 2 (IBRV) 15 0 15 0 0 0 0 T4 1 (BVDV) 15 0 15 0 0 0 0 T5 2 (BVDV) 15 0 15 0 0 0 0 Total Vaccinations (T2, T3, T4 & T5): 59 0 59 0 0 0 0

** All calves were born between 47 and 123 days. The miscarriage occurred between 25 and 43 days.

*** Blood samples prior to lactation were not obtained from one calf in T1 (# 315) and T4 (# 92). T1 calves with pre-lactation antibodies had a titer of> 23 for BVDV-1 (type 1 BVDV) and a titer of> 19 for BVDV-2 (type 2 BVDV). All 58 pre-feeding serum samples collected from T2, T3, T4 and T5 calves were negative (<2) for neutralizing antibodies.

Example 2

생성물 계통의 효능을 평가하기 위한 필드 안전성 연구를 수행하였다. 본 실시예는 78일간 연구의 중간 결과를 요약한 것이다. 본 연구는 번식에 앞서 소 비기관염-바이러스 설사-파라인플루엔자₃, 변형 생 바이러스-캄필로박터 페투스-렙토스피라 카니콜라-그립포티포사-하르조-익테로해모라지애-포모나 박테린 (PregGuard™ FP 9)으로 예방접종시킨 임신 최초 3개월의 교배종 식용 암소에게, 위약 (멸균 수 희석제), 또는 소 비기관염-바이러스 설사-파라인플루엔자₃-호흡기 합포체 변형-생 바이러스 백신/렙토스피라 카니콜라-그립포티포사-하르조-익테로해모라지애-포모나 박테린 (Bovi-Shield

4+L5)의 3가지 몫 중의 1가지를 투여한 것과 관련한 안전성을 비교하였다.BOVI-SHIELD

Field safety studies were conducted to assess the efficacy of the product line. This example summarizes the intermediate results of the 78 day study. This study was carried out prior to breeding for bovine rhinitis-virus diarrhea-parainfluenza ₃ , modified live virus-campylobacter petus-leptospira canicola-gripforty-sa-harzo-acterohaemorazia-formonabacterin (PregGuard ™ To edible cows of the first 3 months of pregnancy vaccinated with FP 9), placebo (sterile water diluent), or bovine rhinitis-virus diarrhea-parainfluenza ₃ -respiratory synaptic modification-live virus vaccine / lepspiracanicola-grip Forty-Phossa-Harzo-Icterohaemorazia-pomona bacterin (Bovi-Shield

The safety associated with administration of one of the three shares of 4 + L5) was compared.

FP 4+L5의 배정 몫의 근육내 용량을 투여하였다. 예방접종 후에 불리한 국소 또는 전신 반응은 전혀 관찰되지 않았다. 이들 동물을 대상으로 하여 매주 건강 상태를 관찰하였다. 302마리 대조군 암소 (T1) 중의 1마리를 개봉하고, Bovi-Shield FP 4+L5로 예방접종시킨 모든 암소 [T2 (n=102), T3 (n=100), 및 T4 (n=95)]는 임신한 것으로 확인되었다. 유산율은 대조군 (T1)과 예방접종한 동물 (T2 내지 T4)에 대해 각각 0.3% 대 0.0%로 극히 유사하였다. 개봉시킨 T1 암소 (#286)로부터의 혈청학적 시험 결과는 상기 분획물 모두에 대한 0일째로부터의 역가에 있어 실질적인 변화가 없었다는 것을 지시해주었다.On day 0, blood samples were collected from each animal and all animals were examined by ultrasound to confirm pregnancy. On day 0, intramuscular doses of placebo were administered to each of the 304 control animals randomly assigned to treatment group T1. To each of the 303 animals assigned to three vaccine treatment groups T2 (n = 106), T3 (n = 101), and T4 (n = 96), Bovi-Shield

An intramuscular dose of an allocation share of FP 4 + L5 was administered. No adverse local or systemic reactions were observed after vaccination. These animals were examined weekly for health. All cows opened in 302 control cows (T1) and vaccinated with Bovi-Shield FP 4 + L5 [T2 (n = 102), T3 (n = 100), and T4 (n = 95)] Was found to be pregnant. Abortion rates were extremely similar at 0.3% versus 0.0% for control (T1) and vaccinated animals (T2 to T4), respectively. Serological test results from the unsealed T1 cow (# 286) indicated no substantial change in titer from day 0 for all of the fractions.

FP 4+L5 백신을 투여한, 미리 예방접종시킨 대략 임신 최초 3개월의 동물의 78일에 걸친 유산율이 위약을 투여한 동물 보다 높지 않았다는 것을 입증해 주었는데, 이는 예방접종한 임신한 동물에서 이들 백신 성분이 안전하다는 것을 뒷받침해준다.This field safety study, Bovi-Shield

The abortion rate over 78 days in approximately the first 3 months of vaccinated animals with the FP 4 + L5 vaccine was not higher than that of placebo-treated animals, which was vaccinated in vaccinated pregnant animals. Support the ingredients in safety.

research plans:

Treatment number Processing description Number of animals Vaccination Check your pregnancy Work Volume Route T1 Sterile Diluents (Placebo) 304 0 2 ml IM 77 days T2 BOVI-SHIELD FP 4 + L5 106 0 2 ml IM 77 days T3 BOVI-SHIELD FP 4 + L5 101 0 2 ml IM 77 days T4 BOVI-SHIELD FP 4 + L5 96 0 2 ml IM 77 days

Processing steps:

Animals were observed weekly for general health. On day 0, animals were observed for general health status, their pregnancy status was confirmed, and two serum separation tubes (SSTs) of blood were collected from each animal. Placebo (sterile diluent) and BOVI-SHIELD FP 4 + L5 vaccine were prepared and 1-dose (2-ml) was administered to the neck side of the appropriate animal via the intramuscular route. Each animal in treatment group T1 received a dose of placebo, while animals in treatment groups T2, T3, and T4 received a share of each dose of BOVI-SHIELD FP 4 + L5. Observation of inappropriate systemic reactions was performed for approximately 1 hour after vaccination.

On day 77, a second pregnancy test was performed on all animals. All non-pregnant animals (opened) were recorded as aborted. Two SST blood samples (acute) were collected from these animals. This sample was processed into serum, labeled and stored frozen. After approximately two weeks, two additional SST blood samples (recovery phases) were collected from the aborted animals, the samples were processed into serum, labeled and stored frozen. On day 0, acute and convalescent serum samples were shipped to a veterinary diagnostic laboratory for bovine abortion serology evaluation.

result:

Table 7: Summary of Dose and Post-Vaccination Responses for Modified Live Virus Components

Table 8: Summary of Abortion Results by Treatment

Table 9: Serological Test Results

Efficacy and Post-Vaccination Response of Investigational Vaccines:

The efficacy of the modified live IBRV, BVDV, PIV ₃ , and BRSV components of each share of BOVI-SHIELD FP 4 + L5 administered on day 0 exceeded the release requirements as detailed in Table 7. All additional tests required for release for commercial distribution were satisfactory. No adverse local or systemic reactions were observed after vaccination on day 0.

Summary of miscarriage results:

As summarized in Table 8, the lactic acid data collected for 77 days were similar for the control and vaccine treatment groups. Only one (0.3%) of 303 control animals (T1) aborted during this study, and none of the 297 animals vaccinated (T2 to T4).

Serological test results for the aborted control cow (T1) (animal number 286) are shown in Table 9.

conclusion:

FP 4+L5 백신을 투여한, 미리 예방접종시킨 임신 최초 3개월 동물의 78일에 걸친 유산율이, 위약을 투여한 동물 보다 높지 않았다는 것을 입증해 주었는데, 이는 예방접종한 임신한 동물에서 이들 백신 성분이 안전하다는 것을 뒷받침해준다.This field safety study, Bovi-Shield

The abortion rate over 78 days in the first three months of pre-vaccinated animals given FP 4 + L5 vaccine was not higher than that of placebo-treated animals, indicating that these vaccine components in vaccinated pregnant animals Support this is safe.

Summary of Efficacy and Post-Vaccination Responses to Modified Live Virus Components Treatment number Day 0 treatment Log ₁₀ TCID ₅₀ Capacity Post-vaccination observations for adverse local or systemic reactions IBRV BVDV PIV ₃ BRSV T1 Sterile Diluent - - - - 0/304 T2 Bovi-4 + L5 6.0 4.2 6.5 4.8 0/106 T3 Bovi-4 + L5 6.0 4.2 6.7 5.0 0/101 T4 Bovi-4 + L5 6.0 4.5 6.5 4.7 0/96

Summary of miscarriage result by treatment Processing group Number of animals Number of miscarriage (ratio) Etiology IBRV Infection BVD infection Congenital defects Unknown T1 (control) 302 1 (0.3%) 0 0 0 One T2 to T4 (vaccinated) 297 0 (0.0%) 0 0 0 0 T2 102 0 (0.0%) 0 0 0 0 T3 100 0 (0.0%) 0 0 0 0 T4 95 0 (0.0%) 0 0 0 0

Serological test results Processing group Work Leptospirogenic Fraction IBR BVD vibrio T1 (# 268) Pomona Canicola Grippo Itero Harzo 0 100-400 Neg-100 Neg-400 Neg-100 Neg-200 16-32 4-8 Neg 77 200 100 100 100 Neg 16 8 Neg 91 200 100 100 100 Neg 32 8 Neg

Example 3

생성물 계통을 대상으로 하여, 필드 안전성 연구를 수행하였다. 본 연구는 번식에 앞서 Bovi-Shield

4+L5로 예방접종시킨 임신 제2기의 홀슈타인 (Holstein) 출산 미경험 암소에게, 위약 (멸균 수 희석제), 또는 소 비기관염-바이러스 설사-파라인플루엔자₃-호흡기 합포체 변형-생 바이러스 백신/렙토스피라 카니콜라-그립포티포사-하르조-익테로해모라지애-포모나 박테린 (Bovi-Shield

4+L5)의 3가지 몫 중의 1가지를 투여한 것과 관련한 안전성을 비교하였다. -1일째에, 각 동물로부터 혈액 샘플을 수집하고, 모든 동물을 직장 촉진 (rectal palpation)함으로써 검사하여 임신 상태를 확인하였다. 0일째에, 무작위로 처리 그룹 T1으로 할당된 238마리의 대조군 동물 각각에 근육내 용량의 위약을 투여하였다. 3가지 백신 처리 그룹 T2 (n=80), T3 (n=79), 및 T4 (n=78)로 할당된 237마리 동물 각각에게, Bovi-Shield

4+L5의 배정 몫의 근육내 용량을 투여하였다. 각 몫의 백신은 방출과 상업적 분배에 대한 모든 시험 요구 조건을 만족시켰다. 예방접종 후에 불리한 국소 또는 전신 반응은 전혀 관찰되지 않았다. 이들 동물을 대상으로 하여 분만을 통한 건강 상태를 매일 관찰하고, 각 신생 송아지의 건강 상태를 결정하였다. 4마리 동물을 본 연구로부터 격리시켰는데, 2마리 (1-T1 및 1-T2)는 상해를 입었고, 2마리 (1-T1 및 1-T4)는 번식일이 부정확하였다. 유산한 동물을 대상으로 하여, 이용 가능한 태아 및 태반 샘플을 동정하고, 예방접종 전 (-1일째)에 급성 및 회복기 혈청을 시험하여 유산의 가능한 원인을 결정하였다. 유산율과 정상적인 분만율은 대조군 (T1)과 예방접종한 동물 (T2 내지 T4)에 대해 매우 유사하였다. 236마리 대조군 동물 중의 11마리 (4.7%)와 예방접종한 (T2 내지 T4) 235마리 동물 중의 14마리 (6.0%)가 본 연구 동안 유산하였다. 25마리 유산아 중의 1마리 (T4)가 IBRV 감염에 기인한 것이었다. 나머지 24마리 유산 증례에 대한 병인론은 네오스포라 (Neospora) 감염, 선천성 결함, 및 공지되지 않은 원인인데, 이는 각각 9마리 (T1=3, T2=1, T3=1, 및 T4=4), 1마리 (T1), 및 14마리 (T1=7, T2=5, 및 T3=2)에게서 일어났다. 건강하게 살아있는 송아지와 분만 도중에 사망한 정상 송아지를 분만한 어미 동물의 비율로 이루어진, 정상적인 전체 분만율은 대조군 (T1) 그룹에 대해서는 95.3% (225/236)이고, 예방접종한 (T2 내지 T4) 그룹에 대해서는 93.6% (220/235)이었다.Bovi-shield

Field safety studies were performed on product lines. This study prior to breeding Bovi-Shield

Holstein in the second trimester of pregnancy vaccinated with 4 + L5, placebo (sterile water diluent), or bovine rhinitis-virus diarrhea-parainfluenza ₃ -respiratory combinatorial strain-live virus vaccine / lepspira Canicola-Grip Potifosa-Harzo-Iterohaemorazia-Pomona Bacterin (Bovi-Shield

The safety associated with administration of one of the three shares of 4 + L5) was compared. On day -1, blood samples were collected from each animal, and all animals were examined by rectal palpation to confirm pregnancy. On day 0, an intramuscular dose of placebo was administered to each of the 238 control animals randomly assigned to treatment group T1. To each of 237 animals assigned to three vaccine treatment groups T2 (n = 80), T3 (n = 79), and T4 (n = 78), Bovi-Shield

An intramuscular dose of an allocation of 4 + L5 was administered. Each portion of the vaccine met all test requirements for release and commercial distribution. No adverse local or systemic reactions were observed after vaccination. These animals were observed daily for their health through childbirth and the health of each newborn calf was determined. Four animals were isolated from this study, with two (1-T1 and 1-T2) injured and two (1-T1 and 1-T4) inaccurate breeding days. For aborted animals, available fetal and placental samples were identified and acute and convalescent serum tested prior to vaccination (day-1) to determine possible causes of abortion. Abortion rates and normal delivery rates were very similar for control (T1) and vaccinated animals (T2 to T4). Eleven (4.7%) of 236 control animals and 14 (6.0%) of 235 animals vaccinated (T2 to T4) aborted during this study. One of the 25 aborted infants (T4) was due to IBRV infection. The etiology for the remaining 24 abortion cases is Neospora infection, congenital defects, and unknown causes, 9 of each (T1 = 3, T2 = 1, T3 = 1, and T4 = 4), 1 Horses (T1), and 14 (T1 = 7, T2 = 5, and T3 = 2). The normal overall delivery rate, which consists of healthy live calves and the mothers who delivered normal calves who died during delivery, was 95.3% (225/236) for the control (T1) group and vaccinated (T2 to T4) groups. For 93.6% (220/235).

4+L5 백신을 투여한, 미리 예방접종시킨 대략 임신 제2기 동물이 유사한 유산율과 정상 송아지 분만율을 나타내었다는 것을 입증해 주었는데, 이는 예방접종한 임신한 동물에서 이들 백신 성분이 안전하다는 것을 뒷받침해준다.The results of this field safety study are either placebo or Bovi-Shield.

It was demonstrated that approximately preterm vaccinated animals given the 4 + L5 vaccine had similar miscarriage rates and normal calf delivery, supporting the safety of these vaccine components in vaccinated pregnant animals. .

research plans:

Treatment number Vaccination Power Before Breeding Number of animals Pregnancy status and days of serum sample collection One intramuscular dose administered in the second trimester of pregnancy Post-Vaccination Monitoring Work Processing description T1 Bovi-Shield 4 + L5 238 -One 0 Sterile Diluents (Placebo) -Observe daily health and miscarriage-Observe new calf health T2 Bovi-Shield 4 + L5 80 -One 0 Bovi-Shield 4 + L5 T3 Bovi-Shield 4 + L5 79 -One 0 Bovi-Shield 4 + L5 T4 Bovi-Shield 4 + L5 78 -One 0 Bovi-Shield 4 + L5

process:

Pre-processing Steps:

Prior to commencing this study, a breeding day for a healthy mating lactating cow with Bovi-Shield 4 + L5 vaccination history was provided prior to breeding. Animals were assigned to treatment groups at random. On day -1, two serum separation tubes (SSTs) of blood were collected from each animal and all animals were examined by rectal palpation to confirm pregnancy. Blood samples were processed into serum, labeled and stored frozen. These animals were assessed daily for general health status and miscarriage at day -1 through delivery.

Processing steps:

On day 0, placebo and Bovi-Shield 4 + L5 vaccine were prepared and administered to the neck side of the appropriate animal via the intramuscular route. Each T1 animal received a dose of placebo (sterile water diluent). Each of the T2, T3 and T4 animals received a dose of BOVI-SHIELD 4 + L5. Observation of harmful systemic reactions was performed for approximately 1 hour after vaccination.

Animals were observed daily for general health status and miscarriage through childbirth. Aborted animals were identified, two SST blood samples (acute) were collected, processed with serum, labeled and stored frozen. Available aborted fetal and placental samples were tested to determine possible causes of abortion. Approximately two weeks after abortion, two additional SST blood samples (recovery phases) were collected from the aborted animals, the samples were processed into serum, labeled and stored frozen.

Childbirth Steps:

The health status of each calf was determined. All calf mortality rates were also assessed to determine the cause of death, with the exception of three egg-related mortality and all fetotomies.

result:

Table 10: Summary of modified live virus components and post-vaccination responses

Table 11: Summary of Abortion Results by Treatment

Table 12: Summary of Delivery Results by Treatment

Investigational vaccines and post-vaccination reactions:

The modified live IBRV, BVDV, PIV ₃ , and BRSV components of each share of Bovi-Shield 4 + L5 administered on day 0 are detailed in Table 10. All additional tests required for release for commercial distribution were satisfactory. No adverse local or systemic reactions were observed after vaccination on day 0.

Summary of miscarriage results:

As summarized in Table 11, abortion results were similar for the control and vaccine treatment groups. Eleven of 236 control animals (T1) (4.7%) and 14 of 235 vaccinated animals (T2 to T4) (6.0%) were aborted during this study. One of the 25 aborted infants (T4) was due to IBRV. The remaining 24 aborted babies are due to neospora, congenital defects, and unknown causes, which are nine (T1 = 3, T2 = 1, T3 = 1, and T4 = 4), one (T1) and It occurred in 14 animals (T1 = 7, T2 = 5, and T3 = 2).

Summary of Calf Birth Results:

Calf delivery results for each treatment group are listed in Table 12. The normal overall delivery rate, which consists of healthy live calves and the mothers who delivered normal calves who died during delivery, was 95.3% (225/236) for the control (T1) group and vaccinated (T2 to T4) groups. For 93.6% (220/235). Within the control (T1) group, 79.7% of the calf were alive and healthy, while 20.3% died during delivery. In comparison, 78.4% of calves delivered by vaccinated animals (T2 to T4) were alive and healthy, and 21.2% of these calves died during the birth process. One T4 animal delivered a dead calf, which was due to neospora infection.

conclusion:

4+L5 백신을 투여한, 미리 예방접종시킨 임신 제2기 동물이 유사한 유산율과 정상 송아지 분만율을 나타내었다는 것을 입증해 주었는데, 이는 예방접종한 임신한 동물에서 이들 백신 성분이 안전하다는 것을 뒷받침해준다.The results of this field safety study are either placebo or Bovi-Shield.

It has been demonstrated that pre-vaccinated second trimester animals given 4 + L5 vaccine showed similar miscarriage rates and normal calf delivery, supporting the safety of these vaccine components in vaccinated pregnant animals.

Summary of Efficacy and Post-Vaccination Responses to Modified Live Virus Components Processing group Day 0 Processing * Log ₁₀ TCID ₅₀ Capacity Post-vaccination observations for adverse local or systemic reactions IBRV BVDV PIV ₃ BRSV T1 Sterile Diluent - - - - 0/238 ** T2 Bovi-4 + L5 6.0 4.2 6.5 4.8 0/80 ** T3 Bovi-4 + L5 6.0 4.2 6.7 5.0 0/79 T4 Bovi-4 + L5 6.0 4.5 6.5 4.7 0/78 **

* The Bovi-4 (Bovi-Shield 4) + L5 shares listed above met all test requirements for release and commercial distribution.

** Animals # 30075 (T2) and Animals # 30400 (T1) were isolated from this study because they were injured on days 110 and 181, respectively. Animal # 19642 (T4) and Animal # 30064 (T1) were isolated from this study on day 238 due to incorrect breeding days. Data for these four animals were excluded from all summaries and analyzes.

Summary of miscarriage result by treatment Processing group Number of animals Number of miscarriage (ratio) Etiology IBRV Neospora infection Congenital defects Unknown T1 (control) 236 11 (4.7%) 0 3 One 7 T2 to T4 (vaccinated) 235 14 (6.0%) One 6 0 7 T2 79 6 (7.6%) 0 One 0 5 T3 79 3 (3.8%) 0 One 0 2 T4 77 5 (6.5%) One 4 0 0

Summary of calf delivery results by treatment Processing group Normal calf delivery rate * (total number of animals / animals giving birth to a normal calf) Normal calf ** Live Healthy Calf Rate Mortality due to difficult birth T1 (control) 95.3% (225/236) 79.7% 20.3% T2 to T4 (vaccinated) 93.6% (220/235) 78.4% 21.2% *** T2 92.4% (73/79) 83.6% 16.4% T3 96.2% (76/79) 76.6% 23.4% T4 92.2% (71/77) 75.0% 23.6% ***

* If the calf is alive or the calf is normal, but due to a difficult birth, the mother animal was determined to deliver a normal calf.

** The ratio includes two sets of twins for T1 and one set of twins for T3.

** The mortality rate for one abnormal calf at T4 was due to neospora infection.

Example 4

FP 4+L5)의 3가지 몫 중의 1가지를 투여한 것과 관련한 안전성을 비교한 필드 안전성 연구를 수행하였다.Bovine rhinitis-virus diarrhea-parainfluenza ₃ , vaccine, modified live virus, Campylobacter petus-reptospira canicola-gripportosa-harzo-ecterohaemorazia-pomonabacterin (PregGuard ™ FP) 9) to approximately 3rd trimester gestational cattle vaccinated with placebo (sterile water diluent), or bovine rhinitis-virus diarrhea-parainfluenza ₃ -respiratory complex alteration-live virus vaccine / leptospira canicola-grippoti Posa-Harzo-Icterohaemoria-pomona bacterin (Bovi-Shield

Field safety studies were performed comparing the safety associated with administration of one of three shares of FP 4 + L5).

FP 4+L5의 배정 몫의 2-ml 용량을 투여하였다. 각 몫의 백신은 방출과 상업적 분배에 대한 모든 시험 요구 조건을 만족시켰다. 예방접종 후에 불리한 국소 또는 전신 반응은 전혀 관찰되지 않았다. 이들 동물을 대상으로 하여 분만을 통한 건강 상태를 관찰하고, 각 신생 송아지의 건강 상태를 결정하였다.Prior to vaccination on day 0, blood samples were collected from each animal and all animals were examined by rectal palpation to confirm pregnancy. Each of the 150 control animals randomly assigned to treatment group T1 received a 2-ml intramuscular dose of sterile diluent (placebo). To each of 198 animals assigned to three vaccine treatment groups T2 (n = 66), T3 (n = 67), and T4 (n = 65), Bovi-Shield

A 2-ml dose of an allotment of FP 4 + L5 was administered. Each portion of the vaccine met all test requirements for release and commercial distribution. No adverse local or systemic reactions were observed after vaccination. These animals were observed for health through childbirth and the health of each newborn calf was determined.

FP 4+L5 백신을 투여한, 미리 예방접종시킨 대략 임신 제3기 동물이 유사한 유산율과 정상 송아지 분만율을 나타내었다는 것을 입증해 주었는데, 이는 예방접종한 임신한 동물에서 이들 백신 성분이 안전하다는 것을 뒷받침해준다.The normal overall delivery rate, consisting of the percentage of healthy live calves and mothers who delivered during delivery (5) or due to harmful climatic conditions (1) normal calf delivery, was 99.3% (148) for the control (T1) group. / 149) and 99.5% (198/196) for the vaccinated (T2 to T4) group. The results of this field safety study are either placebo or Bovi-Shield.

It was demonstrated that approximately preterm vaccinated animals vaccinated with the FP 4 + L5 vaccine had similar miscarriages and normal calf delivery, supporting the safety of these vaccine components in vaccinated pregnant animals. Do it.

research plans:

Treatment number Vaccination Power Before Breeding Number of animals Pregnancy status and days of serum sample collection One intramuscular dose administered to tertiary animals Post-Vaccination Monitoring Work Processing description T1 PregGuard FP 150 0 0 Sterile Diluents (Placebo) -Observe daily health and miscarriage-Observe new calf health T2 PregGuard FP 66 0 0 Bovi-Shield FP 4 + L5 T3 PregGuard FP 67 0 0 Bovi-Shield FP 4 + L5 T4 PregGuard FP 65 0 0 Bovi-Shield FP 4 + L5

process:

Pre-processing Steps:

Prior to starting this study, before propagation, bovine rhinitis-virus diarrhea-parainfluenza ₃ , vaccine, modified live virus, Campylobacter petus-leptospira canicola-gripportifosa-harzo-ecterohaemoria- A breeding day was provided for inexperienced livestock cows of healthy hybrids with a history of vaccination with Pomonabacterin (PregGuard ™ FP 9). Animals were assigned to treatment groups at random. After vaccination, healthy newborn calf health observations were assessed weekly without knowledge of treatment assignments. In addition, laboratory personnel were not able to participate in treatment group assignments.

Processing steps:

On day 0, two serum separation tubes (SSTs) of blood were collected from each animal and all animals were examined by rectal palpation to confirm pregnancy. Blood was processed into serum, labeled and stored frozen. Placebo and Bovi-Shield FP 4 + L5 vaccines were also prepared and administered to the right neck region of the appropriate animal via the intramuscular route. Each T1 animal received a 2-ml dose of placebo (sterile diluent). T2, T3 and T4 animals each received a 2-ml dose of Bovi-Shield FP 4 + L5. Observation of harmful systemic reactions was performed for approximately 1 hour after vaccination.

Animals were observed for general health and miscarriage through childbirth. Aborted animals were identified, two SST blood samples (acute) were collected, processed with serum, labeled and stored frozen. Approximately two weeks after abortion, two additional SST blood samples (recovery phases) were collected from the aborted animals, the samples were processed into serum, labeled and stored frozen.

Childbirth Steps:

The health status of each calf was determined. All abnormal calf (mortality) was due to difficult acid or adverse climatic conditions.

Exclusion / Withdrawal Criteria:

Animal # 71 (T1) and Animal # 288 (T3) were isolated from this study due to the injury observed on Days 0 and 12, respectively. Animal # 237 (T3) delivered calves on day 21 but was isolated from the study on day 22 because the calves could not be placed to determine their health status. Data for these three animals were excluded from all summaries and analyzes.

result:

Table 13: Summary of modified live virus components and post-vaccination responses

Table 14: Summary of miscarriage results by treatment

Table 15: Diagnostic laboratory serological test results for abortion cases

Table 16: Summary of calf delivery results by treatment

Investigational vaccines and post-vaccination reactions:

The titers of modified live IBRV, BVDV, PIV ₃ , and BRSV components of each share of Bovi-Shield FP 4 + L5 administered on day 0 are indicated in Table 13. No adverse local or systemic reactions were observed after vaccination.

Summary of miscarriage results:

As summarized in Table 14, only 1 (0.7%) of 149 control animals (T1) and 1 of 196 animals (T2 to T4) vaccinated were aborted during this study. Diagnostic test results on fetal and / or placental samples did not determine the cause of these abortions. The results of diagnostic serology tests on two abortion cases are presented in Table 15. Leptospira pomona, L. on either animal. Canicola ( L. canicola ), L. L. grippotyphosa (grippo), L. L. icterohaemorrhagiae (ltero), L. There were no substantial changes in acute and recovery levels prior to vaccination of serum antibodies to L. hardjo , IBRV, and BVDV.

Summary of Calf Birth Results:

The normal total delivery rate, consisting of the proportion of healthy live calves and normal calves that died during delivery, is given in Table 16. Normal calf delivery rates were 99.3% (148/149) for the control (T1) group and 99.5% (195/196) for the vaccinated (T2 to T4) groups. All calves, except the six normal calves, were delivered healthy and alive. Five calves died during the birthing process and one died due to adverse weather conditions.

conclusion:

FP 4+L5 백신을 투여한, 미리 예방접종시킨 임신 제3기 동물이 유사한 유산율과 정상 송아지 분만율을 나타내었다는 것을 입증해 주었는데, 이는 예방접종한 임신한 동물에서 이들 백신 성분이 안전하다는 것을 뒷받침해준다.The results of this field safety study are either placebo or Bovi-Shield.

It was demonstrated that pre-vaccinated tertiary gestational animals that received the FP 4 + L5 vaccine had similar abortion rates and normal calf delivery, supporting the safety of these vaccine components in vaccinated pregnant animals. .

Summary of Efficacy and Post-Vaccination Responses to Modified Live Virus Components Processing group Day 0 Processing * Log ₁₀ TCID ₅₀ Capacity Post-vaccination observations for adverse local or systemic reactions IBRV BVDV PIV ₃ BRSV T1 Sterile Diluent - - - - 0 T2 Bovi-Shield 4 (Bovi-4 FP + L5) 6.0 4.2 6.5 4.8 0 T3 Bovi-Shield 4 (Bovi-4 FP + L5) 6.0 4.2 6.7 5.0 0 T4 Bovi-Shield 4 (Bovi-4 FP + L5) 6.0 4.5 6.5 4.7 0

* The Bovi-Shield 4 modified live virus components listed above and the L5 components for each share of Bovi-4 (Bovi-Shield 4) FP + L5 listed above met all test requirements for release and commercial use.

Summary of miscarriage result by treatment Processing group Number of animals Number of miscarriage (ratio) * etiology T1 (control) 149 1 (0.7%) Unknown T2 to T4 (vaccinated) 196 1 (0.5%) Unknown

Animals # 332 (T1) and Animals # 284 (T3) aborted on Day 26 and Day 13, respectively.

Summary of calf delivery results by treatment Processing group Normal calf delivery rate * (animals / total animals giving birth to normal calves) Normal calf Live Healthy Calf death rate T1 (control) 99.3% (148/149) 146 2 T2 to T4 (vaccinated) 99.5% (195/196) 192 ** 4 T2 100.0% (66/66) 66 ** One T3 98.5% (64/65) 63 One T4 100.0% (65/65) 63 2

Mothers were determined to deliver normal calves in the event that the calf is alive or calf is normal, but died due to difficulty in ovulation (5) or climatic conditions (1).

** T2 animals delivered one set of twins.

Claims (15)

A pregnant animal previously vaccinated with a therapeutically effective amount of a vaccine composition comprising a modified bovine herpes virus (BHV-1) is administered to a pregnant animal previously administered a therapeutically effective amount of a vaccine composition comprising a modified bovine herpes virus (BHV-1). A method for preventing the miscarriage of a pregnant animal caused by the BHV-1 virus, comprising. A therapeutically effective amount of a vaccine composition comprising a modified bovine herpes virus (BHV-1) is given to a lactating pregnant animal previously vaccinated with a therapeutically effective amount of a vaccine composition comprising a modified bovine herpes virus (BHV-1). A method for preventing infection of a lactating pregnant animal comprising administering. Modified bovine herpes virus (BHV-1), and bovine viral diarrhea virus (BVDV) to pregnant animals previously vaccinated with a therapeutically effective amount of a vaccine composition comprising modified bovine herpes virus (BHV-1); Modified live parainfluenza virus type 3 (PIV3); Modified bovine respiratory syncytial virus (BRSV); Leptospira car Nicolas (Leptospira canicola), Leptospira grip Fourteen fossa (Leptospira grippotyphosa), Leptospira Borg Peter seniyi Har crude-plastic support teuno (Leptospira borgpetersenii hardjo-prajitno), Leptospira ripening Tero to Mora jiae (Leptospira icterohaemmorrhagiae), Leptospira interrogating elegans Leptospira interrogans pomona , Leptospira borgpetersenii hardjo-bovis , Leptospira Bratislava and Campylobacter fetus among at least one antigen selected from the group consisting of A method for preventing the miscarriage of a pregnant animal caused by BHV-1 virus, comprising administering a therapeutically effective amount of a vaccine composition comprising. Modified bovine herpes virus (BHV-1), and bovine viral diarrhea virus (BVDV) to lactating pregnant animals pre-vaccinated with a therapeutically effective amount of a vaccine composition comprising modified bovine herpes virus (BHV-1) ; Modified live parainfluenza virus type 3 (PIV3); Modified bovine respiratory syncytial virus (BRSV); Leptospira Canicola, Leptospira Grip Potifosa, Leptospira Borghterseniyi Harzo-Prazitno, Leptospira Ichterohaemorazia, Leptospira Interogans pomona, Leptospira Borgpeterceni Bharatlova-Bazillot-Hbartzobis A method for preventing infection of a lactating pregnant animal comprising administering a therapeutically effective amount of a vaccine composition comprising at least one antigen selected from the group consisting of fetus. A pregnant animal previously vaccinated with a therapeutically effective amount of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV) is administered to administer a therapeutically effective amount of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV). A method for preventing the miscarriage of a pregnant animal caused by BVDV virus, comprising. A therapeutically effective amount of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV) is given to a pregnant pregnant woman previously vaccinated with a therapeutically effective amount of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV). A method for preventing infection of a lactating pregnant animal comprising administering. Modified bovine viral diarrhea virus (BVDV), and bovine herpes virus (BHV-1) to pregnant animals pre-vaccinated with a therapeutically effective amount of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV); Modified live parainfluenza virus type 3 (PIV3); Modified bovine respiratory syncytial virus (BRSV); Leptospira Canicola, Leptospira Grip Potifosa, Leptospira Borghterseniyi Harzo-Prazitno, Leptospira Ichterohaemorazia, Leptospira Interogans pomona, Leptospira Borgpeterceni Bharatlova-Bazillot-Hbartzobis A method for preventing abortion of a pregnant animal caused by BVD virus comprising administering a therapeutically effective amount of a vaccine composition comprising at least one antigen selected from the group consisting of fetus. Pre-vaccinated with a therapeutically effective amount of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV), modified bovine viral diarrhea virus (BVDV), and bovine herpes virus (BHV-1); Modified live parainfluenza virus type 3 (PIV3); Modified bovine respiratory syncytial virus (BRSV); Leptospira Canicola, Leptospira Grip Potifosa, Leptospira Borghterseniyi Harzo-Prazitno, Leptospira Ichterohaemorazia, Leptospira Interogans pomona, Leptospira Borgpeterceni Bharatlova-Bazillot-Hbartzobis Therapeutic treatment of a vaccine composition comprising modified bovine viral diarrhea virus (BVDV) in lactating pregnant animals vaccinated prior to breeding in a therapeutically effective amount of a vaccine composition comprising at least one antigen selected from the group consisting of fetuses A method for preventing infection of a lactating pregnant animal, comprising administering an effective amount. In pregnant animals pre-vaccinated with a therapeutically effective amount of a vaccine composition comprising at least one modified bovine herpes virus (BHV-1) and at least one modified bovine viral diarrhea virus, modified live parainfluenza virus type 3 (PIV3) ); Modified bovine respiratory syncytial virus (BRSV); Modified St. 1) and administering a therapeutically effective amount of a vaccine composition comprising bovine viral diarrhea virus. 2. A method for preventing miscarriage of a pregnant animal caused by BHV-1 and BVDV viruses. Modified live parainfluenza virus type 3 to lactating pregnant animals pre-vaccinated with a therapeutically effective amount of a vaccine composition comprising at least one modified bovine herpes virus (BHV-1) and at least one modified bovine viral diarrhea virus. (PIV3); Modified bovine respiratory syncytial virus (BRSV); Leptospira Canicola, Leptospira Grip Potifosa, Leptospira Borgpetersenini Harzo-Prazitno, Leptospira Ichterohaemorazia, Leptospira Interogans pomona, Leptospira Borgpetersenyi Bratislava and St. A method for preventing infection of a lactating pregnant animal comprising administering a therapeutically effective amount of a vaccine composition comprising bovine herpes virus (BHV-1) and bovine viral diarrhea virus. Modified pregnant parainfluenza virus type 3 (PIV3) to pregnant animals previously vaccinated with a therapeutically effective amount of a vaccine composition comprising at least one modified bovine herpes virus (BHV-1) and at least one bovine viral diarrhea virus; Modified bovine respiratory syncytial virus (BRSV); Leptospira Canicola, Leptospira Grip Potifosa, Leptospira Borghterseniyi Harzo-Prazitno, Leptospira Ichterohaemorazia, Leptospira Interogans pomona, Leptospira Borgpeterceni Bharatlova-Bazillot-Hbartzobis Of pregnant animals caused by BHV-1 and BVDV viruses, comprising administering a therapeutically effective amount of a vaccine composition comprising modified live bovine herpes virus (BHV-1) and bovine viral diarrhea virus with Fetus. How to prevent miscarriage. The method of claim 1, wherein the vaccine further comprises an adjuvant. The method of claim 1, wherein the animal is a cow or an inexperienced cow. 9. The method of any one of claims 2, 4, 6 and 8, wherein the animal is a pregnant cow lactating a calf. The method of claim 1, wherein the amount of the vaccine composition administered is about 0.1 to about 5.0 ml per dose.